Lentinan, a Shiitake Mushroom ß-Glucan, Downregulates the Enhanced PD-L1 Expression Induced by Platinum Compounds in Gastric Cancer Cells -


  • Hiroko Ina School of Nursing and Health, Aichi Prefectural University
  • Kenji Ina Department of Psychosomatic Medicine, Nagoya, Japan
  • Megumi Kabeya Department of Pharmacy, Nagoya, Japan
  • Satoshi Kayukawa Department of Clinical Oncology, Nagoya, Japan
  • Takashi Yoshida Department of Clinical Oncology, Nagoya, Japan
  • Masahiko Yoneda School of Nursing and Health, Aichi Prefectural University




Lentinan, gastric cancer, programmed cell death ligand 1, mitogen-activated protein kinase.


 Background: Despite recent therapeutic improvements, the prognosis of unresectable gastric cancer remains poor. Upregulation of programmed cell death ligand 1 (PD-L1) in tumor cells is believed to be an important mechanism to escape from the host immune response. The expression of PD-L1 in tumors is regulated in a highly complex manner by various upstream signaling molecules, depending on the cell type. Given that the efficacy of chemotherapeutic agents for metastatic gastric cancer is limited due to immune escape caused by enhanced PD-L1 expression, PD-1/PD-L1 targeted immunotherapy may be a promising alterative for chemotherapy. However, immune checkpoint inhibitor monotherapy has shown clinical benefits in less than 20% of patients with gastric cancer and its underlying mechanism remains to be elucidated. On the other hand, lentinan, a glucan purified from Shiitake mushrooms, has significant immune-stimulating effects and has been reported to improve survival in patients with metastatic gastric cancer receiving chemotherapy. In the current study we investigated the mechanism by which lentinan increases the chemotherapeutic efficacy by focusing on the expression of PD-L1.

Methods: To evaluate the effects of lentinan as well as antineoplastic agents, the expression of PD-L1 and associated molecules was analyzed by real-time polymerase chain reaction and western blotting using the human gastric cancer cell lines, NUGC3, MKN1, and MKN45.

Results: Treatment with either cisplatin or oxaliplatin dose-dependently enhanced PD-L1 mRNA and protein expression through the mitogen-activated protein kinase (MAPK) pathway in gastric cancer cells. However, lentinan treatment inhibited the platinum drug-stimulated expression of PD-L1 in gastric cancer cells mainly by suppressing MAPK signaling without affecting the phosphatidylinositol-3 kinase/AKT pathway or transcription factors.

Conclusions: Platinum-based drugs enhanced the expression of PD-L1 via the MAPK pathway in gastric cancer cells. Lentinan downregulated PD-L1 expression induced by either cisplatin or oxaliplatin, suggesting that a combination of this glucan and platinum-based chemotherapy could restore the chemosensitivity of cells.


International Agency for Research on Cancer. GLOBOCAN 2018: estimated cancer incidence, mortality and prevalence worldwide. Available online: http://globocan.iarc.fr/ Default.aspx (accessed on 17 June 2020).

Lenz HJ, Lee FC, Haller DG, et al. Extended safety and efficacy data on S-1 plus cisplatin in patients with untreated, advanced gastric carcinoma in a multicenter phase II study. Cancer 2007; 9: 33-40. https://doi.org/10.1002/cncr.22329 DOI: https://doi.org/10.1002/cncr.22329

Koizumi W, Narahara H, Hara T, et al. S-1 plus cisplatin versus S-1 alone for first line treatment of advanced gastric cancer (SPIRITS trial): A phase III trial. Lancet Oncol 2008; 9: 215-221. https://doi.org/10.1016/S1470-2045(08)70035-4 DOI: https://doi.org/10.1016/S1470-2045(08)70035-4

Hironaka S, Sugimoto N, Yamaguchi K, et al. S-1 plus cisplatin leucovorin versus S-1 plus leucovorin and oxaliplatin versus S-1 plus cisplatin in patients with advanced gastric cancer: a randomized, multicenter, open-label, phase 2 trial. Lancet Oncol 2016; 17: 99-108. https://doi.org/10.1016/S1470-2045(15)00410-6 DOI: https://doi.org/10.1016/S1470-2045(15)00410-6

Ina K, Hirade K, Kabeya M, et al. Long-term survivors of metastatic gastric cancer for >5 years after chemotherapy initiation. Cancer Reports and Reviews 2019; 3: 1-5. https://doi.org/10.15761/CRR.1000183 DOI: https://doi.org/10.15761/CRR.1000183

Chen DS, Mellman I. Oncology meets immunology: the cancer-immunity. Immunity 2013; 39: 1-10. https://doi.org/10.1016/j.immuni.2013.07.012 DOI: https://doi.org/10.1016/j.immuni.2013.07.012

Topalian SL, Drake CG, Pardoll DM. Targeting the PD-1/ B7-H1 (PD-L1) pathway to activate anti-tumor immunity. Curr Opin Immunol 2012; 24: 207-2012. https://doi.org/10.1016/j.coi.2011.12.009 DOI: https://doi.org/10.1016/j.coi.2011.12.009

Pardoll DM. The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer 2012; 12: 252-264. https://doi.org/10.1038/nrc3239 DOI: https://doi.org/10.1038/nrc3239

Taube JM, Klein A, Brahmer JR, et al. Association of PD-1, PD-1 ligands, and other features of the tumor immune microenvironment with response to anti-PD-1 therapy. Clin Cancer Res 2014; 20: 5064-5074. https://doi.org/10.1158/1078-0432.CCR-13-3271 DOI: https://doi.org/10.1158/1078-0432.CCR-13-3271

Thompson ED, Zahurk M, Murphy A, et al. Patterns of PD-L1 expression and CD8 T cell infiltration in gastric adenocarcinomas and associated immune stroma. Gut 2016. https://doi.org/10.1136/gutjnl-2015-310839 DOI: https://doi.org/10.1136/gutjnl-2015-310839

Tumeh PC, Harview CL, Yearly JH, et al. PD-L1 blockade induces responses by inhibiting adaptive immune resistance. Nature 2014; 515: 568-571. https://doi.org/10.1038/nature13954 DOI: https://doi.org/10.1038/nature13954

Concha-Benavente F, Srivastava RM, Trivedi S, et al. Identification of the Cell-Intrinsic and -Extrinsic Pathways Downstream of EGFR and IFNγ That Induce PD-L1 Expression in Head and Neck Cancer. Cancer Res 2016; 76: 1031-43. https://doi.org/10.1158/0008-5472.CAN-15-2001 DOI: https://doi.org/10.1158/0008-5472.CAN-15-2001

Jiang X, Zhou J, Giobbie-Hurder A, Wargo J, Hodi FS. The association of MAPK in melanoma cells resistant to BRAF inhibition promotes PD-L1 expression that is reversible by MEK and PI3K inhibition. Clin Cancer Res 2013 19; 598-609. https://doi.org/10.1158/1078-0432.CCR-12-2731 DOI: https://doi.org/10.1158/1078-0432.CCR-12-2731

Stutvoet T, Kol A, de Vries EG, et al. MAPK pathway activity plays a key role. J Pathol 2019; 249: 52-64. https://doi.org/10.1002/path.5280 DOI: https://doi.org/10.1002/path.5280

Parsa AT, Waldron JS, Panner A, et al. Loss of tumor suppressor PTEN function increases B7-H1 expression and immunoresistance in glioma. Nat Med 2007; 13: 84-88. https://doi.org/10.1038/nm1517 DOI: https://doi.org/10.1038/nm1517

Wolfle SJ, Strebovsky J, Bartz H, et al. PD-L1 expression on tolerogenic APCs is controlled by STAT-3. Eur J Immunol 2011; 41: 413-424. https://doi.org/10.1002/eji.201040979 DOI: https://doi.org/10.1002/eji.201040979

Bouillez A, Rajabi H, Jin C, et al. MUC-1 integrates PD-L1 induction with repression of immune effectors in non-small-cell lung cancer. Oncogene 2017; 36: 4037-46. https://doi.org/10.1038/onc.2017.47 DOI: https://doi.org/10.1038/onc.2017.47

Latchman Y, Wood CR, Chernova T, et al. PD-L2 is a second ligand for PD-1 and inhibits T cell activation. Nat Immunol 2001; 2: 261-8. https://doi.org/10.1038/85330 DOI: https://doi.org/10.1038/85330

Schmid P, Hegde PS, Zou W, et al. Association of PD-L2 expression in human tumors with atezolizumab activity. J Clin Oncol 2016; 34(Suppl 15): 11506. https://doi.org/10.1200/JCO.2016.34.15_suppl.11506 DOI: https://doi.org/10.1200/JCO.2016.34.15_suppl.11506

Yearly JH, Gibson C, Yu N, et al. PD-L2 expression in human tumors: Relevance to anti-PD-1 therapy in cancer. Clin Cancer Res 2017; 23: 3158-67. https://doi.org/10.1158/1078-0432.CCR-16-1761 DOI: https://doi.org/10.1158/1078-0432.CCR-16-1761

Chihara G, Hamuro J, Maeda Y, Arai Y, Fukuoka F. Fractionation and purification of the polysaccharides with marked antitumor activity, especially lentinan, from Lentinus edodes (Berk.) Sing Cancer Res 1970; 30: 2776-2781.

Ina K, Kataoka T, Ando T. The use of lentinan for treating gastric cancer. Anticancer Agents Med Chem 2013; 13: 681-8. https://doi.org/10.2174/1871520611313050002 DOI: https://doi.org/10.2174/1871520611313050002

Oba K, Kobayashi M, Matsui T, Kodera Y, Sakamoto J. Individual patient based meta-analysis of lentinan for unresectable/recurrent gastric cancer. Anticancer Res 2009; 29: 2739-46.

Wang H, Cai Y, Zheng Y, Bai Q, Xie D, Yu J. Efficacy of biological response modifier lentinan with chemotherapy for advanced cancer: a meta-analysis. Cancer Med 2017; 6: 2222-33. https://doi.org/10.1002/cam4.1156 DOI: https://doi.org/10.1002/cam4.1156

Ren L, Perera C, Hemar Y. Antitumor activity of mushroom polysaccharides: a review. Food Funct 2012; 3: 1118-30. https://doi.org/10.1039/c2fo10279j DOI: https://doi.org/10.1039/c2fo10279j

Aleem, E. β-glucans and their applications in cancer therapy: focus on human studies. Anticancer Agents Med Chem 2013; 13: 709-19. https://doi.org/10.2174/1871520611313050005 DOI: https://doi.org/10.2174/1871520611313050005

Yoshino S, Nishikawa K, Morita S, et al. Randomised phase III study of S-1 alone versus S-1 plus lentinan for unresectable or recurrent gastric cancer. Eur J Cancer 2016; 65: 164-71. https://doi.org/10.1016/j.ejca.2016.06.012 DOI: https://doi.org/10.1016/j.ejca.2016.06.012

Higashi D, Seki K, Ishibashi Y, et al. The effect of lentinan combination therapy for unresectable advanced gastric cancer. Anticancer Res 2012; 32: 2365-8.

Ina H, Yoneda M, Kanda M, et al. Lentinan, a shiitake mushroom β-glucan, stimulates tumor-specific adaptive immunity through PD-L1 down-regulation in gastric cancer cells. Med Chem (Los Angeles) 2016; 6: 710-4. https://doi.org/10.4172/2161-0444.1000419 DOI: https://doi.org/10.4172/2161-0444.1000419

Schefe JH, Lehmann KE, Buchsmann IR, Unger T, Funke-Kaiser H. Quantitative real-time RT-PCR data analysis: current concepts and the novel gene expression’s CT difference formula. J Mol Med 2006; 84: 901-10. https://doi.org/10.1007/s00109-006-0097-6 DOI: https://doi.org/10.1007/s00109-006-0097-6

Inoue Y, Yoneda M, Zhao J, et al. Molecular cloning and characterization of chick SPACRCAN. J Biol Chem 2006; 281: 10381-8. https://doi.org/10.1074/jbc.M508161200 DOI: https://doi.org/10.1074/jbc.M508161200

Yamada H, Yoneda M, Inaguma S, et al. Infliximab counteracts tumor necrosis factor-α-enhanced induction of matrix metalloproteinases that degrade claudin and occludin in non-pigmented ciliary epithelium. Biochem Pharmacol 2013; 85: 1770-82. https://doi.org/10.1016/j.bcp.2013.04.006 DOI: https://doi.org/10.1016/j.bcp.2013.04.006

Kanda Y. Investigation of the freely-available easy-to-use software “EZR” (Easy R) for medical statistics. Bone Marrow Transplant 2013; 48: 452-8. https://doi.org/10.1038/bmt.2012.244 DOI: https://doi.org/10.1038/bmt.2012.244

Grabosch S, Bulatovic M, Zeng F, et al. Cisplatin-induced immune modulation in ovarian cancer mouse models with distinct inflammation profiles. Oncogene 2018; 38: 2380-93. https://doi.org/10.1038/s41388-018-0581-9 DOI: https://doi.org/10.1038/s41388-018-0581-9

Fournel L, Wu Z, Stadler N, et al. Cisplatin increases PD-L1 expression and optimizes immune check-point blockade in non-small lung cancer. Cancer Letters 2019; 464: 5-14. https://doi.org/10.1016/j.canlet.2019.08.005 DOI: https://doi.org/10.1016/j.canlet.2019.08.005

Paz-Ares L, Luft A, Vicente D, et al. Pembrolizumab plus chemotherapy for squamous non–small-cell lung cancer. N Engl J Med 2018; 379; 2040-51. https://doi.org/10.1056/NEJMoa1810865 DOI: https://doi.org/10.1056/NEJMoa1810865

Gadgeel S, Rodoriguez-Abreu D, Speranza G, et al. Updated analysis from KEYNOTE-189: Pembrolizumab or placebo plus pemetrexed an previously untreated metastatic nonsquamous non-small cell lung cancer. J Clin Oncol 2020; JCO1903136. https://doi.org/10.1200/JCO.19.03136 DOI: https://doi.org/10.1200/JCO.19.03136

Reck M, Rodríguez-Abreu D, Robinson AG, et al. Pembrolizumab versus chemotherapy for PD-L1-positive non-small-cell lung cancer. N Engl J Med 2016; 375: 1823-33. https://doi.org/10.1056/NEJMoa1606774 DOI: https://doi.org/10.1056/NEJMoa1606774

Chen J, Jiang CC, Jin L, Zhang XD. Regulation of PD-L1: A novel role of pro-survival signaling in cancer. Ann Oncol 2016; 27: 409-16. https://doi.org/10.1093/annonc/mdv615 DOI: https://doi.org/10.1093/annonc/mdv615

Sun C, Mezzadra R, Schumacher T. Regulation and function of the PD-L1 checkpoint. Immunity 2018; 48: 434-52. https://doi.org/10.1016/j.immuni.2018.03.014 DOI: https://doi.org/10.1016/j.immuni.2018.03.014

Gowrishanker K, Gunatilake D, Gallagher SJ, Tiffen J, Rizos H, Hersey P. Inducible but not constitutive expression of PD-L1 in human melanoma cells is dependent on activation of NF-β. PLoS One 2015; 10. https://doi.org/10.1371/journal.pone.0123410 DOI: https://doi.org/10.1371/journal.pone.0123410

Vetvicka V, Vetvickova J. Anti-infectious and anti-tumor activities of β–glucans. Anticancer Res 2020; 40: 3139-45. https://doi.org/10.21873/anticanres.14295 DOI: https://doi.org/10.21873/anticanres.14295

Xu X, Pan C, Zhang L, Ashida H. Immunomodulatoryβ-glucan from Lentinus edodes activates mitogen-activated protein kinases and nuclear factor-β in murine RAW 264.7 macrophages. J Biol Chem 2011; 286: 31194-8. https://doi.org/10.1074/jbc.M111.246470 DOI: https://doi.org/10.1074/jbc.M111.246470

Chihara G, Maeda Y, Hamuro J, et al. Inhibition of mouse sarcoma 180 by polysaccharides from Lentinus edodes (Berk.) Sing. Nature 1969; 222: 687-8. https://doi.org/10.1038/222687a0 DOI: https://doi.org/10.1038/222687a0

Ina K, Furuta R. Image of Month: Complete response of metastatic gastric cancer to chemo-immunotherapy. Indian J Med Res 2017; 146: 141. https://doi.org/10.4103/ijmr.IJMR_132_16 DOI: https://doi.org/10.4103/ijmr.IJMR_132_16




How to Cite

Hiroko Ina, Kenji Ina, Megumi Kabeya, Satoshi Kayukawa, Takashi Yoshida, & Masahiko Yoneda. (2020). Lentinan, a Shiitake Mushroom ß-Glucan, Downregulates the Enhanced PD-L1 Expression Induced by Platinum Compounds in Gastric Cancer Cells -. Journal of Analytical Oncology, 9, 1–10. https://doi.org/10.30683/1927-7229.2020.09.01